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Abstract
The present study was carried out to (1) evaluate the effect of sheep oocyte quality, hormonal and serum additives, media type and concentration of epidermal growth factor (EGF) on in vitro maturation (IVM) of sheep oocytes. (2) investigate the effect of ram breed and type of media on in vitro capacitation of rams spermatozoa (3) study the effect of type of medium, type of serum and EGF concentration on in vitro fertilization (IVF) and embryonic development. Sheep ovaries were collected from local slaughterhouse. Aspirated oocytes were cultured in TCM-199 or SOF for 24-26 h at 39°C under 5% CO2 in air and 95% humidity. The present results indicated that supplementing TCM-199 with 10IU /ml FSH + 10 IU/ml LH achieved the highest percentage of MII oocytes (40.36 %). Adding EES to the maturation media (SOF or TCM-199) significantly (P<0.001) increased the percentage oocytes with complete cumulus cells expansion (82.93 % and 83.33%, respectively). Hormonal additives and serum (EES or FCS) in the maturation medium, obviously improved the rate of oocytes either class A or B developed to MII. Adding 40 ng /ml EGF achieved the highest percentage of oocytes that reached MII. The highest percentage of live sperm with reacted acrosome (L2) (true acrosome reaction) was performed by Rahmani breed followed by Farafra and Barki rams especially when sperm were incubated in B.O. medium. Adding EES to TCM-199 or SOF achieved the highest percentages of fertilization and embryonic development compared with supplementing FCS.